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Genome-wide RNA analysis of the effects of MASTL silencing in breast cancer cells

ABSTRACT: Genome-wide RNA analysis of the effects of MASTL silencing in breast cancer cells

PROVIDER: PRJNA545096 | ENA |

REPOSITORIES: ENA

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Genome-wide RNA analysis of the effects of MASTL silencing in breast cancer cells

Project description:The effect of short term (24h and 48h) silencing of MASTL for total transcriptome was studied in human breast cancer cell line (MDA-MB-231). Microtubule-associated serine/threonine-protein kinase-like (MASTL) is a mitosis-accelerating kinase with emerging roles in cancer progression. However, possible cell cycle–independent mechanisms behind its oncogenicity remain ambiguous. Here, we identify MASTL as an activator of cell contractility and MRTF-A/SRF (myocardin-related transcription factor A/serum response factor) signaling. Depletion of MASTL increased cell spreading while reducing contractile actin stress fibers in normal and breast cancer cells and strongly impairing breast cancer cell motility and invasion. Transcriptome and proteome profiling revealed MASTL-regulated genes implicated in cell movement and actomyosin contraction, including Rho guanine nucleotide exchange factor 2 (GEF-H1, ARHGEF2) and MRTF-A target genes tropomyosin 4.2 (TPM4), vinculin (VCL), and nonmuscle myosin IIB (NM-2B, MYH10). Mechanistically, MASTL associated with MRTF-A and increased its nuclear retention and transcriptional activity. Importantly, MASTL kinase activity was not required for regulation of cell spreading or MRTF-A/SRF transcriptional activity. Taken together, we present a previously unknown kinase-independent role for MASTL as a regulator of cell adhesion, contractility, and MRTF-A/SRF activity.

2020-04-14 | GSE131833 | GEO

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FAM83H-AS1 silencing in MCF7 breast cancer cells

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Transgelin silencing induces different processes in differrent breast cancer cell lines

Project description:Transgelin is a protein which has been described as a marker of several cancers. Interestingly, the literature describes both up- and down-regulation of transgelin in the tumor in comparison with non-tumor tissue. The mechanisms of transgelin function in cancer are considerably unknown. Transgelin is abundant especially in smooth muscle cells. It is associated with actin stress fibers. These contractile structures participate among others in cell motility, adhesion or morphology. We focused on transgelin function in breast cancer. Initially, we studied transgelin role in cell migration in the breast cancer cell lines BT 549 and PMC 42. Using xCELLigence system we reached opposite results in two cell lines. Transgelin silencing increased and decreased migration of PMC 42 and BT 549 cells, respectively. To further clarify these contradictory results, we performed an experiment to quantify proteomic changes after transgelin silencing in these two cell lines using quantitative proteomics (iTRAQ-2DLC-MS/MS). Our results confirmed transgelin function in the migration of BT 549 cells and suggested transgelin role in apoptosis and small molecule biochemistry in PMC 42 cells.

2020-02-21 | PXD016316 | Pride

FAM83H-AS1 silencing in MCF7 breast cancer cells

Project description:We silenced FAM83H-AS1 shRNAs in cell line MCF7 carried a ~75% silencing compared to thenegative control (NC). We evaluated the role of FAM83H-AS1 on oncogenic phenotypes in the MCF7 breast cancer cell line model. The knockdown of FAM83H-AS1 was achieved with ~75% of silencing efficiency. A complete transcriptomic analysis after silencing of FAM83H-AS1 revealed an impact on the global expression.

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Transcriptome analysis of QPRT silencing in breast cancer cells

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Silencing ferrochelatase in breast cancer cells MDA-MB-231

Project description:Silencing ferrochelatase in breast cancer cells MDA-MB-231

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Genome-wide Scan for Methylation Profiles in Breast Cancer

Project description:Genome wide DNA methylation profiling of normal and breast cancer samples. The Illumina Infinium 450k Human DNA methylation Beadchip v1.1 was used to obtain DNA methylation profiles across approximately 485577 CpGs in breast cancer and normal samples. Samples included 40 normal, 80 breast cancer samples.

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Proteomic analysis of the secretome from 4T1 breast cancer cells upon Stat3 silencing.

Project description:Stat3 has been acknowledged as an oncogene that has dual protumorigenic activity in a cell-intrinsic way, by promoting cancer cell proliferation and survival and in a paracrine mode, acting as suppressor of immune cell attack. We demonstrated that Stat3 silencing with a small interfering RNA (siRNA) induced a senescence program in Stat3-addicted cancer cells and leads to the production of a senescence associated secretory phenotype (SASP) that has several antitumor properties. The composition of the SASP induced by Stat3 silencing is poorly characterized. The goal of the project was to analyze the protein content in the secretome released by the mouse breast cancer cell line 4T1 after Stat3 silencing with a small interfering RNA.

2020-01-21 | PXD013948 | Pride

Genome-wide gene expression analysis of MDA-MB-468 human breast cancer cells treated with PLA2G7 shRNA

Project description:The aim was to investigate the functional role of PLA2G7 in breast cancer using PLA2G7 silencing in cultured breast cancer cells. For stable knockdown of PLA2G7, shRNAs were transduced to MDA-MB-468 cells before genome-wide gene expression analysis. Gene expression profiles of the PLA2G7 silenced samples were compared with the scrambled control treated samples and two replicate samples were studied for each treatment.

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